AMPA受体参与的出生后大鼠海马发育早期的电生理学特点
陈雪怡¹, 张皑峰², 赵文^1,3,4, 高钰丹^1,3,4, 段红梅^1,3,4, 郝鹏^1,3,4, 杨朝阳^1,3,4, 李晓光^1,3,4,*
1首都医科大学神经生物学系，北京 100069；²首都医科大学附属北京友谊医院口腔科，北京 100050；³北京航空航天大学生物与医学工程学院生物材料和神经再生北京市实验室，北京 100191；⁴北京航空航天大学大数据精准医疗高精尖创新中心，北京 100191
摘要
本研究旨在探讨α-氨基-3-羟基-5-甲基-4-异恶唑丙酸(AMPA)受体参与的出生后大鼠海马发育早期的电生理学特点。选择出生后0.5月龄、1月龄、2月龄和3月龄Wistar大鼠共计48只(每组各12只)。应用全细胞膜片钳技术及MED64平面微电极阵列技术检测海马CA1区锥体神经元的被动膜特性及AMPA受体参与的自发兴奋性突触后电流(spontaneous exctitatory postsynaptic current, sEPSC)和场兴奋性突触后电位(field excitatory postsynaptic potential, fEPSP)。结果显示，海马CA1区锥体神经元在出生后0.5~3月龄期间，在被动膜特性方面表现为：膜电容与静息膜电位无显著性变化；膜输入电阻与时间常数均显著下降。在主动膜特性方面，呈现出阶段性变化：0.5~1月龄期间，sEPSC的反应表现为：振幅显著升高，频率明显增大，上升时间及下降时间显著增加；1~3月龄期间，sEPSC的反应特性与0.5~1月龄期间相反。此外，0.5~3月龄期间，海马CA1区诱发出的fEPSP范围明显扩大，而幅值显著减小；各月龄海马CA1区诱发出的fEPSP幅值均可被AMPA受体竞争性拮抗剂6-氰基-7-硝基喹喔啉-2,3-二酮(CNQX)明显降低。以上结果提示，在出生后大鼠海马发育早期过程中，AMPA受体作为调节突触传递和突触联系的主要兴奋性受体，可以促进海马的发育及功能成熟。
关键词： AMPA受体; 出生后发育早期; 全细胞膜片钳技术; MED64平面微电极阵列记录技术
分类号：R742

Electrophysiological characteristics of hippocampal postnatal early development mediated by AMPA receptors in rats
CHEN Xue-Yi¹, ZHANG Ai-Feng², Zhao Wen^1,3,4, GAO Yu-Dan^1,3,4, DUAN Hong-Mei^1,3,4, HAO Peng^1,3,4, ANG Zhao-Yang^1,3,4, LI Xiao-Guang^1,3,4,*
1Department of Neurobiology, Capital Medical University, Beijing 100069, China；²Department of Stomatology, Beijing Friendship Hospital, Capital Medical University, Beijing 100050, China；³Beijing Key Laboratory for Biomaterials and Neural Regeneration, School of Biological Science and Medical Engineering, Beihang University, Beijing 100191, China；⁴Beijing Advanced Innovation Center for Big Data-Based Precision Medicine, Beihang University, Beijing 100191, China
Abstract
The present study was aimed to investigate the electrophysiological characteristics of hippocampal postnatal early development mediated by α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptors in rats. Forty-eight Wistar rats were divided into postnatal 0.5-, 1-, 2- and 3-month groups (n = 12). Spontaneous excitatory postsynaptic currents (sEPSCs) and field excitatory postsynaptic potentials (fEPSPs) mediated by AMPA receptors were recorded to evaluate the changes in the intrinsic membrane properties of hippocampal CA1 pyramidal neurons by using patch-clamp and MED64 planar microelectrode array technique respectively. The results showed that, during the period of postnatal 0.5–3 months, some of the intrinsic membrane properties of hippocampal CA1 pyramidal neurons, such as the membrane capacitance (Cm) and the resting membrane potential (RMP), showed no significant changes, while the membrane input resistance (Rin) and the time constant (τ) of the cells were decreased significantly. The amplitude, frequency and kinetics (both rise and decay times) of sEPSCs were significantly increased during the period of postnatal 0.5–1 month, but they were all decreased during the period of postnatal 1–3 months. In addition, the range of evoked fEPSPs in hippocamal CA1 region was significantly expanded, but the fEPSP amplitudes were decreased significantly during the period of postnatal 0.5–3 months. Furthermore, the evoked fEPSPs could be significantly inhibited by extracellular application of the AMPA receptor antagonist 6-cyano- 7-nitroquinoxaline-2,3-dione (CNQX). These results suggest that AMPA receptor may act as a major type of excitatory receptor to regulate synaptic transmission and connections during the early stage of hippocampal postnatal development, which promotes the development and functional maturation of hippocampus in rats.


Key words: AMPA receptor; early postnatal development; patch clamp technique; planar microelectrode array recording technique

收稿日期：2017-12-23　　录用日期：2018-03-30
通讯作者：李晓光　　E-mail: lxgchina@sina.com
DOI: 10.13294/j.aps.2018.0031
引用本文：
陈雪怡, 张皑峰, 赵文, 高钰丹, 段红梅, 郝鹏, 杨朝阳, 李晓光. AMPA受体参与的出生后大鼠海马发育早期的电生理学特点[J]. 生理学报 2018; 70 (2): 106-114.
CHEN Xue-Yi, ZHANG Ai-Feng, Zhao Wen, GAO Yu-Dan, DUAN Hong-Mei, HAO Peng, ANG Zhao-Yang, LI Xiao-Guang. Electrophysiological characteristics of hippocampal postnatal early development mediated by AMPA receptors in rats. Acta Physiol Sin 2018; 70 (2): 106-114 (in Chinese with English abstract).



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